Optical rotations were measured on a DIP-370 polarimeter (JASCO, Oklahoma, OK, USA) at 589 nm using a sample cell of path length 0.1 dm. UV spectra were obtained on a Cary 50 Bio UV-Visible spectrophotometer (Varian, Palo Alto, CA, USA). IR spectra of the compounds, as thin films on CaF2 discs, were recorded using an IRAffinity-1 Fourier Transform spectrophotometer (Shimadzu, Kyoto, Japan). NMR spectra were acquired on an Inova 500 MHz spectrometer (Varian, Palo Alto, CA, USA) operating at 500 (1H) or 125 (13C) MHz using the residual solvent signals as internal references (CD3OD δH 3.30, δC 49.0; DMSO-d6 δH 2.50, δC 39.5; CDCl3 δH 7.26, δC 77.0). Samples were placed in 3-mm Shigemi NMR tubes as necessary. HRMS data were obtained from 6210 LC/TOF and 6545 LC/QTOF instruments (Agilent, Santa Clara, CA, USA) using electrospray ionization in positive and negative modes. ESIMS data were also obtained using a 6410 QQQ instrument (Agilent, Santa Clara, CA, USA). Samples were purified using a combination of chromatographic techniques, including high performance liquid chromatography. Purity of the compounds was assessed from their UV absorptions at appropriate wavelengths, or from their 1H NMR spectra. Percent yields of natural products were based on the amount of freeze-dried biological material.
6545 lc q tof
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 6545 LC/Q-TOF is a high-performance liquid chromatography-quadrupole time-of-flight (LC/Q-TOF) mass spectrometer. It is designed for accurate mass measurement and high-resolution analysis of complex samples.
Automatically generated - may contain errors
Lab products found in correlation
Cary 50 bio uv visible spectrophotometer, by Agilent Technologies (1 mentions)
6210 lc tof, by Agilent Technologies (1 mentions)
Inova 500 mhz spectrometer, by Agilent Technologies (1 mentions)
Dip 370 polarimeter, by Jasco (1 mentions)
Eclipse plus c18 rapid resolution hd column, by Agilent Technologies (1 mentions)
Masshunter metlin metabolomics database, by Agilent Technologies (1 mentions)
Masshunter qualitative analysis software, by Agilent Technologies (1 mentions)
1290 infinity 2 lc, by Agilent Technologies (1 mentions)
Advance 3 hd 400 mhz, by Bruker (1 mentions)
Vario el cube, by Elementar (1 mentions)
Eclipse plus c18 rrhd, by Agilent Technologies (1 mentions)
1290 infinity 2, by Agilent Technologies (1 mentions)
Eclipse plus c18 rapid resolution hd, by Agilent Technologies (1 mentions)
7 protocols using 6545 lc q tof
1
Analytical Characterization of Natural Products
2
Phytochemical Profiling of B. ceiba Stamen
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The phytochemical screening of the orange B. ceiba stamen extracts was analyzed using LC-MS with a ZORBAX Eclipse Plus C18 Rapid Resolution HD column (2.1 × 150 mm, 1.8 μm), utilizing a Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry (LC-QTOF-MS) instrument (6545 LC/Q-TOF, Agilent Technology, Santa Clara, CA, USA), which consisted of a binary pump. The samples were dissolved in 70% methanol at a concentration of 1 mg/mL and passed through a 0.22 µm filter. The temperature was maintained at 35 °C, and the injection volume was 1 μL. Elution was performed for 21 minutes with a mobile-phase gradient system starting with 20% ACN and 80% water (0.1% formic acid), with a flow rate of 0.2 mL/min. The MS conditions involved an electrospray ionization (ESI) probe in the negative mode. MassHunter Qualitative Analysis Software (Agilent MassHunter Workstation Qualitative Analysis version 10.0, CA, USA) was used to evaluate the ion’s match with the database, and this was carried out by comparing the measured mass to the exact mass of the molecular formula and the predicted isotope pattern, utilizing the find-by-formula search of the Agilent MassHunter METLIN Metabolomics Database. The chemical characteristics of the compounds were obtained using the Medlin library [60 (link)].
3
Optimized AAV Characterization Protocol
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All protocols documented were optimized from testing different parameters such as AAV titers, sample incubation in different acidic conditions of formic acid (FA) and TFA, times of sample incubation, varying the LC gradients for optimal elution of the AAV VP proteins at the desired of ∼3 min, column temperature, sample injection volume, and different serotypes, with reproducibility and robustness.
Agilent 1290 Infinity II LC: auto-sampler with 20-μL loop for both intact and digested analyses.
Agilent 6545 LC/QTOF: Dual-jet stream source. SeeTable 3 for LC setting details, and Table 4 for MS setting details.
Agilent 1290 Infinity II LC: auto-sampler with 20-μL loop for both intact and digested analyses.
Agilent 6545 LC/QTOF: Dual-jet stream source. See
4
Comprehensive Crude Oil Characterization
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Composition features of the target crude oil were characterized
by the tandem gas chromatography mass spectrometer (7890B-5977A, Agilent,
USA) and electrospray mass spectrometer (6545 LC/Q-TOF, Agilent, USA).
Composition features of the petroleum coke samples were characterized
by elemental analysis (Vario EL Cube, made by Elementar in Germany)
and 13C NMR (Advance III HD 400 MHz, Bruker, Switzerland).
by the tandem gas chromatography mass spectrometer (7890B-5977A, Agilent,
USA) and electrospray mass spectrometer (6545 LC/Q-TOF, Agilent, USA).
Composition features of the petroleum coke samples were characterized
by elemental analysis (Vario EL Cube, made by Elementar in Germany)
and 13C NMR (Advance III HD 400 MHz, Bruker, Switzerland).
5
Metabolite Extraction and Identification
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The fermentation broth of mutants was extracted by EtOAc, and the organic solvent was combined and concentrated in vacuo to obtain crude extract. The extraction was redissolved in MeOH and filtered using 0.22 μm organic membranes to inject into HPLC analysis at a flow rate of 1 mL/min. The solvent gradient system for HPLC was 0–5 min 100–80% A, 5–35 min 80–40% A, 35–50 min 40% A, 50–55 min 40–0% A, 55–60 min 0–100% A, and 60–65 min 100% A (A: 5% ACN/H2O with 0.05% formic acid, B: 100% ACN with 0.05% formic acid). The LC-HRMS analysis was carried out on an Agilent C18 column (Eclipse Plus C18 RRHD, 2.1 × 50 mm). The positive mode electrospray ionization was performed with a linear gradient of 10–100% acetonitrile in ddH2O with 0.1% formic acid over 6.5 min, followed by 100% acetonitrile for 1.5 min with a flow rate of 0.6 mL/min. The LC-HRMS spectra were recorded on an Agilent 1290 Infinity II coupled to 6545 LC/Q-TOF.
6
Identification of Flavonoid Aglycones by LC-MS
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Identification of aglycones after enzymatic hydrolysis by snailase was carried out by a 1290 Infinity II LC System equipped with a ZORBAX Eclipse Plus C18 Rapid Resolution HD (1.8 μm, 2.1 × 150 mm) connected to a G7117C diode array detector and subsequently followed by a 6545 LC/Q-TOF (Agilent, USA) with a multimode ion source. The flow rate was set to 0.3 ml/min, and the column oven was operated at 35°C.
The same mobile phase as for HPLC analysis was used with the following gradients, namely, 1 min at 5 vol% (B), 4 min from 5 to 15 vol% (B), 6 min to 53 vol% (B), 4 min to 100 vol% (B), 6.5 min at 100 vol% (B), 0.5 min to 5 vol%, and post run 5 min at 5 vol% (B). The wavelength during analysis was 290 nm. All solvents were of LC-MS grade.
Mass spectrums were recorded in ESI-negative mode between m/z 100 and 1,000 Da (scan rate: 2 spectra/s) after a 5 min solvent cut. The mass spectrometer was operated as follows: gas temp.: 350°C, gas flow: 10 l/min, nebulizer: 40 psig, vaporizer: 220°C, VCharge: 1.75 kV, VCap: 1 kV, fragmentor: 180 V, and skimmer: 75 V.
Flavonoid aglycones were identified according to the retention time, substance-specific UV absorbance, and the accurate mass of the molecular ion. All identified flavonoid aglycones and relevant data per plant extract by mass spectrometric analysis are presented inSupplementary Table S23 .
The same mobile phase as for HPLC analysis was used with the following gradients, namely, 1 min at 5 vol% (B), 4 min from 5 to 15 vol% (B), 6 min to 53 vol% (B), 4 min to 100 vol% (B), 6.5 min at 100 vol% (B), 0.5 min to 5 vol%, and post run 5 min at 5 vol% (B). The wavelength during analysis was 290 nm. All solvents were of LC-MS grade.
Mass spectrums were recorded in ESI-negative mode between m/z 100 and 1,000 Da (scan rate: 2 spectra/s) after a 5 min solvent cut. The mass spectrometer was operated as follows: gas temp.: 350°C, gas flow: 10 l/min, nebulizer: 40 psig, vaporizer: 220°C, VCharge: 1.75 kV, VCap: 1 kV, fragmentor: 180 V, and skimmer: 75 V.
Flavonoid aglycones were identified according to the retention time, substance-specific UV absorbance, and the accurate mass of the molecular ion. All identified flavonoid aglycones and relevant data per plant extract by mass spectrometric analysis are presented in
7
Kinetics of ISG15 Deconjugation by USP18
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1 μM recombinant hUSP18 (WT, C64S, C64S/C65S, I60N) and 5 μM recombinant pro-hISG15 were incubated for 3 or 10 min at 37°C in the buffer of 25 mM HEPES pH 8.0, 200 mM NaCl, 1 mM TCEP. Cleavage of pro-hISG15(AA1-165) to mature hISG15(AA1-157) was monitored by SDS-PAGE as well as mass spectrometry using 6545 LC/Q-TOF (Agilent technologies) as described above in “recombinant protein expression and purification ”.
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